BIOPESTICIDES REGISTRATION ACTION DOCUMENT

Helicoverpa zea ABA Nucleopolyhedrovirus - U

Pesticide Chemical (PC) Code: 107200

U.S. Environmental Protection Agency

Office of Pesticide Programs

Biopesticides and Pollution Prevention Division

DRAFT – February 4, 2014
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TABLE OF CONTENTS

I. EXECUTIVE SUMMARY...........................................................................................................................4

II. ACTIVE INGREDIENT OVERVIEW .......................................................................................................6

III. REGULATORY BACKGROUND ..............................................................................................................7

A. Applications for Pesticide Product Registration ............................................................................7

B. Food Tolerance Exemption ..............................................................................................................7

IV. RISK ASSESSMENT....................................................................................................................................7

A. Product Analysis Assessment (40 CFR § 158.2120) .......................................................................8

B. Human Health Assessment (40 CFR § 158.2140)...........................................................................8

C. Environmental Assessment (40 CFR § 158.2150).........................................................................14

V. ENVIRONMENTAL JUSTICE.................................................................................................................16

VI. RISK MANAGEMENT DECISION .........................................................................................................17

VII. ACTIONS REQUIRED OF THE REGISTRANT...................................................................................17

VIII. GLOSSARY OF ACRONYMS AND ABBREVIATIONS ......................................................................18

IX. BIBLIOGRAPHY .......................................................................................................................................19

A. Studies Submitted to Support the Active Ingredient, , Helicoverpa zea Nucleopolyhedrovirus 

and Pesticide Product, Heligen, Registration.........................................................................................................19

B. Environmental Protection Agency Risk Assessment Memoranda .............................................21

C. Other References..........................................................................................................................212

APPENDIX A. MICROBIAL PESTICIDES DATA REQUIREMENTS ...........................................................23

APPENDIX B. PESTICIDE PRODUCT................................................................................................................27
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BIOPESTICIDES REGISTRATION ACTION DOCUMENT TEAM

Office of Pesticide Programs (OPP)

Biopesticides and Pollution Prevention Division

Microbial Pesticides Branch

Science Reviews

Ibrahim S. Barsoum, Ph.D. Product Analysis, Human Health

John L. Kough, Ph.D. Product Analysis, Human Health

Shannon Borges Environmental Effects

Regulations

Kimberly Nesci Chief, Microbial Pesticides Branch

Michael Glikes Regulatory Action Leader
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I. EXECUTIVE SUMMARY

Background

In September 2012, MacIntosh & Associates, Inc., on behalf of AgBiTech Pty Ltd., an 

Australian company, submitted applications for a new manufacturing use product (MP), 

Helicoverpa zea ABA Nucleopolyhedrovirus- U (EPA File Symbol 87978-R), and a new enduse product (EP), Heligen (EPA File Symbol 87978-E), to the United States Environmental 

Protection Agency (EPA) under section 3 of the Federal Insecticide, Fungicide, and Rodenticide 

Act (FIFRA).

Helicoverpa zea Nucleopolyhedrovirus (Hz NPV) is a naturally occurring insect virus, originally 

isolated from Helothine Lepidoptera, specifically the cotton bollworm (Helicoverpa zea) and 

tobacco budworm (Heliothis virescens). In 1975, the EPA first registered Heliothis zea NPV 

under the brand name Elcar for use on cotton and tobacco to control the cotton bollworm and 

tobacco budworm. In 1984, the EPA issued a registration standard for the use of Heliothis zea 

NPV on a variety of agricultural crops, and in 1990 the Agency issued a Registration Eligibility 

Decision (RED), which determined that the pesticide poses no unreasonable adverse effect to 

human health and the environment. Systematic entomologists changed the genus from 

"Heliothis" to "Helicoverpa" in the late 1990s.

The proposed (MP) containing this active ingredient is an isolate of the registered strain, and it is 

called Helicoverpa zea ABA Nucleopolyhedrovirus-U (Hz ABA-NPV-U). The proposed EP

containing this active ingredient is Heligen. AgBiTech’s first Hz NPV EP was commercialized in 

Australia under the name ViVUS. The proposed EP is for use in the United Sates in field 

applications to control corn earworm, cotton bollworm, tomato fruitworm, and tobacco budworm 

on food and non-food crops, including sorghum, cereal grains, alfalfa, oilseed, potatoes, pulses, 

cotton, sweet corn, berry fruits, brassica, leafy vegetables, legume vegetables, pome fruits, 

celery, cucurbits, and ornamentals. It is proposed for application by ground or aerial spray or by 

chemigation.

EPA scientists reviewed product analysis, toxicology, and nontarget organism data and 

information (40 CFR §§ 158.2120, 158.2140, and 158.2150, respectively) submitted to support 

the registration of the proposed pesticide products. They found that, overall, such data and 

information are adequate for risk assessment purposes, fulfill the current microbial pesticide data

requirements, and support registration of the proposed product under FIFRA section 3(c)(5).

Product Analysis

For the purposes of FIFRA section 3(c)(5) registration, the product analysis data requirements 

for Helicoverpa zea ABA Nucleopolyhedrovirus-U (Hz ABA-NPV-U),which is both the 

technical grade active ingredient (TGAI) and the MP, and the EP, Heligen, including product 

chemistry and composition, analysis of samples, physical and chemical characteristics, and 

additional data concerning genetic stability, were fulfilled by acceptable guideline studies.

Toxicology

The applicant submitted adequate mammalian toxicology data and information to support the 

proposed pesticide products. Acute oral, pulmonary, and injection toxicity/pathogenicity studies
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showed that, at a single high dose, Hz ABA-NPV-U is not toxic and/or pathogenic via these

routes of exposure. Moreover, the applicant reported that no hypersensitivity incidents occurred

during research, development, or testing of Hz ABA-NPV-U. In light of the results of the acute 

toxicity/pathogenicity data and the absence of hypersensitivity incidents, testing at higher tiers 

(i.e., Tiers II and III) was not required.

Tolerance Exemption

Because Helicoverpa zea Nucleopolyhedrovirus is already exempt from this requirement, the 

applicant did not petition the EPA for an exemption from the requirement of a tolerance for 

residues of the new isolate, Hz ABA-NPV-U. In 1980 the EPA granted an exemption from the 

requirement of a tolerance for residues of the active ingredient Hz NPV in or on all agricultural 

commodities (7 CFR 180.1027) as long as the manufacturer meets certain specifications related 

to identity, contamination control, and safety. In its application to the EPA, the applicant 

provided data and other information to support coverage by the existing exemption from the 

tolerance requirement. Hence, the EPA concludes that there is reasonable certainty that no harm 

will result to the U.S. population, including infants and children, from aggregate exposure to 

residues of Hz ABA-NPV-U. Data and information submitted by the applicant support the 

conclusion that no dietary risks are expected to occur as a result of the use of Hz ABA-NPV-U as 

an active ingredient in pesticide products. The EPA concludes that there will not be significant 

exposure to Hz ABA-NPV-U through food and drinking water. Based on the review of acute oral 

toxicity and pathogenicity data by EPA scientists, the Agency determined that, even if Hz ABANPV-U were present on food or in drinking water, human exposure to this insect virus would not 

result in unreasonable adverse effects.

Occupational Exposure

Despite the low toxicological profile of residues of the active ingredient, Hz ABA-NPV-U, in or 

on all agricultural commodities, the EPA is requiring baseline personal protective equipment 

(PPE) for handlers that, due to their occupation, may be subject to prolonged or repeated 

exposure to the active ingredient. The label on Heligen, the proposed end-use product, directs 

handlers working with Hz NPV in agricultural settings to wear a long-sleeved shirt, long pants, 

socks, and shoes. For future products, EPA may require additional PPE (e.g., protective eyewear

and/or a dust/mist filtering respirator meeting National Institute for Occupational Safety and 

Health (NIOSH) standards of at least N-95, R-95, or P-95) on a product-specific basis. 

Nontarget Organisms

The applicant submitted rationale bridging studies previously submitted for a different isolate of 

Helicoverpa zea Nucleopolyhedrovirus to support the Agency’s waiving nontarget organism 

testing for the new active ingredient (AI). The EPA determined that this new AI is substantially 

similar to the previously registered AI and that previously submitted data would provide an 

adequate measure of hazard for the new AI. Hence, the Agency accepted the bridging rationale

and determined that the data are sufficient to fulfill the relevant microbial pesticide data 

requirements and for risk assessment purposes. Further testing of nontarget organisms at higher 

tier levels (i.e., Tiers II, III, and IV) is not required. The EPA performed an environmental risk 

assessment, based on data and other information provided by the applicant and previously 

submitted data cited by the previous applicant, and determined that adverse effects to nontarget 

organisms are not anticipated from the proposed pesticidal uses of Heligen, which contains the 
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active ingredient Hz ABA-NPV-U. Moreover, the EPA made a “No Effect” determination for 

direct and indirect effects to federally listed threatened and endangered species and their 

designated critical habitats resulting from these same proposed pesticidal uses. 

Public Participation

On October 1, 2009, the EPA announced a new policy to provide a more meaningful opportunity 

for the public to participate in major registration decisions before they occur. According to this 

policy, the EPA intends to provide a public comment period prior to making a registration 

decision for, at minimum, the following types of applications: new active ingredients; first food 

uses; first outdoor uses; first residential uses; or any other registration actions for which the 

Agency believes there may be significant public interest.

 

Consistent with the policy of making registration actions more transparent, the proposed 

pesticide product containing Hz ABA NPV-U, is subject to a 15-day comment period. In 

addition to containing a new active ingredient, the registration of the pesticide product would 

result in the first outdoor and food uses for Hz NPV. The docket identification number associated 

with these registration actions, and accessed through either http://www.regulations.gov or 

http://www.epa.gov/pesticides/regulating/registration-status.html, is EPA-HQ-OPP-2013-0099. 

The following documents are available for comment in EPA-HQ-OPP-2013-0099: (1) draft 

Helicoverpa zea ABA Nucleopolyhedrovirus - U Registration Action Document (BRAD); (2) 

environmental risk assessment for Hz ABA NPV-U; (3) draft label for the manufacturing-use 

product (File Symbol 87978-R), and (4) draft label for the end-use pesticide product, Heligen 

(EPA File Symbol 87978-E). While a final decision on registration is contingent upon review 

and consideration of public comments, EPA believes that, based upon the data and information 

submitted in support of Hz ABA NPV-U and Heligen, it is appropriate to issue the registration.

The basis for this preliminary decision can be found in the risk assessment for Hz NPV, which is 

characterized throughout this BRAD. 

II. ACTIVE INGREDIENT OVERVIEW

Biological Name: Helicoverpa zea ABA Nucleopolyhedrovirus - U

Culture Deposit: American Type Culture Collection: SD-6742 

OPP Chemical Code: 107200

Type of Pesticide: Microbial Pesticide – Insecticide

See Appendix B for specific information (e.g.,

use sites, application rates, methods of application, 

formulation types, and target pests) regarding

the proposed pesticide products containing this 

active ingredient.
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III. REGULATORY BACKGROUND

A. Applications for Pesticide Product Registration

In September 2012, MacIntosh & Associates, Inc. (address: 1203 Hartford Avenue, Saint Paul, 

MN 55116) submitted an application on behalf of AgBiTech Pty. Ltd. (address: P.O. Box 537, 

Richmond, New South Wales 2753) to register a manufacturing-use pesticide product,

Helicoverpa zea ABA Nucleopolyhedrovirus - U (EPA File Symbol 87978-R), and an end-use 

product, Heligen (EPA File Symbol 87978-E), under FIFRA section 3. On April 19, 2013 (76 FR 

23558), EPA announced receipt of the application to register a pesticide product containing a 

new active ingredient for use in agriculture (outdoors). EPA opened a 30-day public comment 

period pursuant to the provisions of FIFRA section 3(c)(4). No comments were received 

following this publication. 

B. Food Tolerance Exemption

Under Federal Food, Drug, and Cosmetic Act (FFDCA) section 408(d), the EPA in 1980 granted 

an exemption from the requirement of a tolerance for residues of the active ingredient 

Helicoverpa zea Nucleopolyhedrovirus in or on all agricultural commodities (40 CFR 180.1027) 

as long as the manufacturer meets certain specifications related to identity, contamination 

control, and safety. In its application to the EPA in 2012, MacIntosh & Associations, on behalf 

of the Australian company AgBiTech, provided data and other information to support coverage 

by the existing exemption from the requirement of a tolerance for residues of Hz ABA NPV-U in 

or on all agricultural commodities. 

IV. RISK ASSESSMENT

In the Federal Register of October 26, 2007 (72 FR 61002), the EPA issued a Final Rule on the 

data requirements to support registration of microbial pesticides and updated the definition for 

microbial pesticides. The rule became effective on December 26, 2007. The data and information 

evaluated for this BRAD were considered in light of these requirements.

The EPA classifies each data submission with an indication of the usefulness of the information 

contained in the documents for risk assessment. A rating of “acceptable” indicates the study is 

scientifically sound and is useful for risk assessment. A “supplemental” rating indicates the data 

provide some information that can be useful for risk assessment. The studies may have certain 

aspects determined not to be scientifically acceptable (“supplemental: upgradeable”). If a study 

is rated as “supplemental: upgradeable,” EPA always provides an indication of what is lacking or 

what can be provided to change the rating to “acceptable.” If there is simply a “supplemental” 

rating, the reviewer will often state that the study is not required by 40 CFR Part 158. Both 

“acceptable” and “supplemental” studies may be used in the risk assessment process as 

appropriate. An “unacceptable” rating indicates that new data must be submitted.

For the acute toxicity data requirements, Toxicity Categories are assigned based on the 

hazard(s) identified from studies and/or other information submitted to the EPA in support of a 

pesticide registration. The EPA classifies the active ingredient or particular product into Toxicity 

Category I, II, III, or IV, where Toxicity Category I indicates the highest toxicity and Toxicity 

Category IV indicates the lowest toxicity (see 40 CFR § 156.62). 
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A. Product Analysis Assessment (40 CFR § 158.2120)

For purposes of registration under FIFRA section 3(c)(5), the EPA determined that all product 

analysis data requirements for Helicoverpa zea ABA Nucleopolyhedrovirus - U have been 

fulfilled. Refer to Tables 1, 2, 3, and 4 in Appendix A for a summary of the data requirements, 

including both generic and product-specific information.

B. Human Health Assessment (40 CFR § 158.2140)

1. Toxicity

All toxicology data requirements for Helicoverpa zea ABA Nucleopolyhedrovirus – U have been 

fulfilled. Acceptable Tier I mammalian toxicology data and information support registration of 

the pesticide product, Heligen. Furthermore, Tier II and Tier III studies were not required for Hz

ABA NPV-U because of the lack of acute toxicity/pathogenicity in the Tier I studies.

For a summary of the toxicology data for Hz ABA NPV-U described in sections IV(B)(1)(a) and 

IV(B)(1)(b) (see pages 8–10), refer to Table 5 in Appendix A. 

a. Acute Toxicity/Pathogenicity – Tier I

The applicant evaluated the identity or similarity between NPV strains by extracting DNA, 

treating with restriction endonuclease and using electrophoresis to compare the gel patterns

(MRID No. 490114-01). The products, AgBiTech's Heligen and the previously registered 

product containing Hz NPV, Gemstar (EPA Registration Number 70051-45), were examined by 

light microscopy using Giemsa stain and X I000 magnification under phase contrast. This 

process confirmed that the products contain a nucleopolyhedrovirus with occlusion bodies 

visually similar to each other. Researchers extracted the virus from both products, and they 

extracted DNA from the virus occlusion bodies. Then, this DNA was digested with restriction 

endonuclease (RENs) and the profile under electrophoresis on agarose was compared with that 

of NPV in Hz NPV (Gemstar). This analysis shows that the Hz ABA NPV-U in the AgBiTech 

product is almost identical to the Hz NPV found in Gemstar. Based on this, data previously 

submitted to support the registration of HzNPV (Gemstar) can be bridged to this new strain.

Acute Oral Toxicity/Pathogenicity (Harmonized Guideline 885.3050; Master Record 

Identification Number (MRID No.489417-06): The EPA rated this scientific rationale for waiver 

request as acceptable for risk assessment purposes (U.S. EPA 2013; Barsoum and Kough). Hz

NPV is a baculovirus with a long history of safe use as a microbial pesticide in the United States. 

Toxicology results from baculoviruses have raised no concerns for human or animal health. 

Baculoviruses have narrow host ranges, and the mode of action is well understood and unique to 

insect hosts, with virtually no adverse effects in people, wildlife, or non-target insects. Based on 

literature search for the period 1980-2012, there are no data demonstrating any toxic effects on 

humans or other mammals exposed to Hz NPV.

Acute Pulmonary Toxicity/Pathogenicity(Harmonized Guideline 885.3150; MRID No.489417-

06): The EPA rated this scientific rationale for waiver request as acceptable for risk assessment 

purposes (U.S. EPA 2013; Barsoum and Kough). Hz NPV is a baculovirus with a long history of 

safe use as a microbial pesticide in the United States. Toxicology results from baculoviruses 

have raised no concerns for human or animal health. Baculoviruses have narrow host ranges, and 
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the mode of action is well understood and unique to insect hosts, with virtually no adverse 

effects in people, wildlife, or non-target insects. Based on literature search for the period 1980-

2012, there are no data demonstrating any toxic effects on humans or other mammals exposed to 

Hz NPV. 

Acute Injection Toxicity/Pathogenicity (Intravenous) (Harmonized Guideline 885.3200; MRID 

No 489417-06 ): The EPA rated this scientific rationale for waiver request as acceptable for risk 

assessment purposes (U.S. EPA 2013; Barsoum and Kough). Hz NPV is a baculovirus with a 

long history of safe use as a microbial pesticide in the United States. Toxicology results from 

baculoviruses have raised no concerns for human or animal health. Baculoviruses have narrow 

host ranges, and the mode of action is well understood and unique to insect hosts, with virtually 

no adverse effects in people, wildlife, or non-target insects. Based on literature search for the 

period 1980-2012, there are no data demonstrating any toxic effects on humans or other 

mammals exposed to Hz NPV.

Hypersensitivity Incidents (Harmonized Guideline 885.3400; MRID 489417-06): The applicant 

reported that no hypersensitivity incidents, including immediate-type or delayed-type reactions 

of humans and domestic animals, occurred during research, development, or testing of Hz ABA 

NPV-U. Any future hypersensitivity incidents must be reported to EPA (refer to test note #3 of 

40 CFR § 158.2140(d)).

Cell Culture (Harmonized Guideline 885.3500) MRID 48941807: Researchers tested for 

infectivity the following cells: human lung fibroblast WI38, primary human dermal fibroblast, 

primate continuous line CV-1, and primary Syrian hamster embryo cells. They incubated the 

four cells with Hz NPV (>1x10 virus particles), commercial Hz NPV, a NPV control, and 

inactivated Hz NPV virus, sub-cultured at days 7 and 14, and examined daily through day 7 and 

at days 14 and 21 after infection for cytopathic effect (CPE). Using Dot Blot techniques, the 

researchers examined the cell cultures for the presence of viral nucleic acid at days 1, 2, 5, 7, 14 

and 21, and then they resuspended the cultures in an appropriate volume based on the number of 

cells collected. Triplicate 25-μL samples were applied to positively charged nylon membrane at 

300 to 22,250 cells per spot for most samples (inactivated virus samples had 0 to 4,250 cells per 

spot). To detect Hz NPV nucleic acids in the tested cells, researchers used purified enzyme 

restriction digested genomic viral DNA from the commercial virus Hz NPV. Helicoverpa zea

cells infected with the commercial virus Hz NPV were used as positive control on the dot blot. 

The researchers used mock-infected H. zea cells (cells exposed to only cell culture media) as a 

negative control on the dot blot, photographed the blots, and stored samples below -60°C. They 

determined that the number of colonies per plate observed after 21 or 25 days for all cell lines 

with the virus infected cells was equivalent to the number of colonies per plate with no 

treatment. They observed no cytopathic effects or replication of Hz NPV exposed to any of the 

mammalian cell lines. Since no Hz NPV nucleic acid was detected, no further testing was 

necessary. The EPA rated this study as acceptable.

Acute Oral Toxicity (Harmonized Guideline 870.1100; Master Record Identification Number 

(MRID No. 489417-06 ): The EPA rated this scientific rationale for waiver request as acceptable 

for risk assessment purposes (U.S. EPA 2013; Barsoum and Kough). Hz NPV is a baculovirus 

with a long history of safe use as a microbial pesticide in the United States. Toxicology results 

from baculoviruses have raised no concerns for human or animal health. Baculoviruses have 

narrow host ranges, and the mode of action is well understood and unique to insect hosts, with 

virtually no adverse effects in people, wildlife, or non-target insects. Based on literature search 
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for the period 1980-2012, there are no data demonstrating any toxic effects on humans or other 

mammals exposed to Hz NPV. The EPA classified the acute oral toxicity/pathogenicity as 

Toxicity Category IV.

Acute Inhalation Toxicity (Harmonized Guideline 870-1300; MRID No. 489417-06 ): The EPA 

rated this scientific rationale for waiver request as acceptable for risk assessment purposes (U.S. 

EPA 2013; Barsoum and Kough). Hz NPV is a baculovirus with a long history of safe use as a 

microbial pesticide in the United States. Toxicology results from baculoviruses have raised no 

concerns for human or animal health. Baculoviruses have narrow host ranges, and the mode of 

action is well understood and unique to insect hosts, with virtually no adverse effects in people, 

wildlife, or non-target insects. Based on literature search for the period 1980-2012, there are no 

data demonstrating any toxic effects on humans or other mammals exposed to Hz NPV. The 

EPA classified the acute inhalation toxicity as Toxicity Category IV.

Acute Dermal Toxicity - Rat (Harmonized Guideline 870.1200; MRID No489417-06 ): The EPA 

rated this scientific rationale for waiver request as acceptable for risk assessment purposes (U.S. 

EPA 2013; Barsoum and Kough). Hz NPV is a baculovirus with a long history of safe use as a 

microbial pesticide in the United States. Toxicology results from baculoviruses have raised no 

concerns for human or animal health. Baculoviruses have narrow host ranges, and the mode of 

action is well understood and unique to insect hosts, with virtually no adverse effects in people, 

wildlife, or non-target insects. Based on literature search for the period 1980-2012, there are no 

data demonstrating any toxic effects on humans or other mammals exposed to Hz NPV. The 

EPA classified the dermal toxicity as Toxicity Category IV.

Acute Eye Irritation Study (Harmonized Guideline 870.2400; MRID No 489417-06 ): The EPA 

rated this scientific rationale for waiver request as acceptable for risk assessment purposes (U.S. 

EPA 2013; Barsoum and Kough). Hz NPV is a baculovirus with a long history of safe use as a 

microbial pesticide in the United States. Toxicology results from baculoviruses have raised no 

concerns for human or animal health. Baculoviruses have narrow host ranges, and the mode of 

action is well understood and unique to insect hosts, with virtually no adverse effects in people, 

wildlife, or non-target insects. Based on literature search for the period 1980-2012, there are no 

data demonstrating any toxic effects on humans or other mammals exposed to Hz NPV. The 

EPA classified the eye irritation as Toxicity Category IV.

Acute Dermal Irritation Study (Harmonized Guideline 870.2500; MRID No489417-06 ): The 

EPA rated this scientific rationale for waiver request as acceptable for risk assessment purposes 

(U.S. EPA 2013; Barsoum and Kough). Hz NPV is a baculovirus with a long history of safe use 

as a microbial pesticide in the United States. Toxicology results from baculoviruses have raised 

no concerns for human or animal health. Baculoviruses have narrow host ranges, and the mode 

of action is well understood and unique to insect hosts, with virtually no adverse effects in 

people, wildlife, or non-target insects. Based on literature search for the period 1980-2012, there 

are no data demonstrating any toxic effects on humans or other mammals exposed to Hz NPV. 

The EPA classified the primary dermal irritation as Toxicity Category IV.

b. Acute Toxicology and Subchronic Toxicity/Pathogenicity – Tier II; 

ReproductiveFertility Effects, Carcinogenicity, Immunotoxicity, and 

Infectivity/Pathogenicity Analysis – Tier III

Tier II and Tier III studies were not required for Helicoverpa zea ABA Nucleopolyhedrovirus -
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U based on the lack of acute toxicity/pathogenicity in the Tier I data requirements.

c. Endocrine Disruptors

As required by the Administrator under the Federal Food, Drug, and Cosmetic Act (FFDCA), the 

EPA has developed the Endocrine Disruptor Screening Program (EDSP) and has begun to 

implement the screening program that is to be used to test all pesticides to determine whether 

certain substances (including pesticide active and other ingredients) may have an effect in 

humans or wildlife similar to an effect produced by a “naturally occurring estrogen, or other such 

endocrine effects as the Administrator may designate.” FFDCA section 408(p)(4) authorizes the 

Administrator, by order, to exempt from the requirements of the Endocrine Disruptor Screening 

Program a biologic substance or other substance if a determination is made that the substance is 

anticipated to not produce any effect in humans similar to an effect produced by a naturally 

occurring estrogenic substance. Between October 2009 and February 2010, EPA issued test 

orders/data call-ins for the first group of 67 chemicals, which contains 58 pesticide active 

ingredients and 9 inert ingredients. Helicoverpa zea Nucleopolyhedrovirus (Hz NPV) is not 

among the group of 58 pesticide active ingredients on the initial list to be screened under the 

EDSP. Hz ABA NPV-U is substantially similar to Hz NPV.

The EPA believes that Hz ABA NPV-U is a substance that would not produce any effect in 

humans similar to an effect produced by a naturally occurring estrogenic substance. As such, and 

pursuant to Section 408(p)(4), the EPA will determine in the future whether it can exempt Hz

ABA NPV-U from the requirements of the Section 408(p)(4) EDSP. In the event the EPA does 

determine to exempt this substance from the EDSP, an order will be issued. For further 

information on the status of the EDSP, the policies and procedures, the list of 67 chemicals, the 

test guidelines and the Tier 1 screening battery, please visit our website: 

http://www.epa.gov/endo/.

2. Federal Food, Drug, and Cosmetic Act (FFDCA) Considerations

Section 408(c)(2)(A)(i) of FFDCA allows the EPA to establish an exemption from the 

requirement for a tolerance (the legal limit for a pesticide chemical residue in or on a food) only 

if the EPA determines that the exemption is “safe.” Section 408(c)(2)(A)(ii) of FFDCA defines 

“safe” to mean that “there is a reasonable certainty that no harm will result from aggregate 

exposure to the pesticide chemical residue, including all anticipated dietary exposures and all 

other exposures for which there is reliable information.” This includes exposure through drinking 

water and in residential settings, but it does not include occupational exposure. Pursuant to 

section 408(c)(2)(B) of FFDCA, in establishing or maintaining in effect an exemption from the 

requirement of a tolerance, the EPA must take into account the factors set forth in section 

408(b)(2)(C) of FFDCA, which require the EPA to give special consideration to exposure of 

infants and children to the pesticide chemical residue in establishing a tolerance exemption, and 

to “ensure that there is a reasonable certainty that no harm will result to infants and children from 

aggregate exposure to the pesticide chemical residue....” Additionally, section 408(b)(2)(D) of 

FFDCA requires that the EPA consider “available information concerning the cumulative effects 

of [a particular pesticide's] . . . residues and other substances that have a common mechanism of 

toxicity.”

The EPA performs a number of analyses to determine the risks from aggregate exposure to 

pesticide residues. First, the Agency determines the toxicity of a pesticide. Second, the EPA 
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examines exposure to the pesticide through food, drinking water, and through other exposures 

that occur as a result of pesticide use in residential settings.

Consistent with section 408(b)(2)(D) of FFDCA, the EPA has reviewed the available scientific 

data and other relevant information, and considered its validity, completeness, and reliability and 

the relationship of this information to human risk. The Agency also considered available 

information concerning the variability of the sensitivities of major identifiable subgroups of 

consumers, including infants and children.

Based on the acute toxicity/pathogenicity data and information discussed previously and 

presented in Table 5 in Appendix A, the data required for a FFDCA risk assessment for 

Helicoverpa zea ABA Nucleopolyhedrovirus - U have been fulfilled.

a. Aggregate Exposure

In examining aggregate exposure, section 408 of FFDCA directs EPA to consider 

available information concerning exposures from the pesticide residue in food and all other 

nonoccupational exposures, including drinking water from ground water or surface water and 

exposure through pesticide use in gardens, lawns, or buildings (residential and other indoor 

uses).

Food Exposure and Risk Characterization: The EPA found that dietary exposure to Helicoverpa 

zea ABA Nucleopolyhedrovirus - U, a naturally occurring insect virus, is anticipated to be 

negligible. Due to the ubiquitous nature of the organism and the concentrations of Hz NPV and 

other closely related viruses that already exist in the environment, the likelihood of increased risk 

to humans or animals from the use of Heligen is low. The EPA concluded that the risk posed to 

adults, infants, and children is likely to be minimal because of the low acute oral 

toxicity/pathogenicity potential of Hz ABA NPV-U and any potential exposure to Hz ABA 

NPV-U also would be minimal.

Drinking Water Exposure and Risk Characterization: Exposure to residues of Helicoverpa zea 

ABANucleopolyhedrovirus - U in consumed drinking water is unlikely. The proposed use 

patterns for Heligen are soil directed and/or soil incorporated, thereby limiting contact with 

surface water by drift and runoff. Furthermore, ground water is not expected to have significant 

exposure to Hz NPV since, like other microorganisms, this microbial pesticide would likely be 

filtered out by the particulate nature of many soil types. If it were to be transferred to surface or 

ground waters that are intended for eventual human consumption (e.g., through spray drift or 

runoff) and directed to wastewater treatment systems or drinking water facilities, it likely would 

not survive the conditions water is subjected to in such systems or facilities, including 

chlorination, pH adjustments, filtration, and/or occasionally high temperatures (Centers for 

Disease Control and Prevention 2009; U.S. EPA 2004). In the remote likelihood that Hz NPV is 

present in drinking water (e.g., water not subject to treatment systems or facilities), its target pest 

specificity and available data indicate no toxicity and/or pathogenicity is likely to occur with any

drinking water exposure to Hz ABA NPV-U that results from pesticide applications made in 

accordance with good agricultural practices (see section IV(B)(1)(a) on pages 8–10 and Table 5

in Appendix A).

Non-occupational, Residential Risk Characterization: Given that Hz ABA NPV-U is naturallyoccurring and usually not stable outside of its insect host, EPA determined that non-occupational 

exposure to the bacterium is unlikely. Repeated exposures to the microorganism from pesticidal 
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applications do not exceed EPA’s level of concern, particularly in light of available data that 

demonstrate Hz ABA NPV-U is not toxic (acute dermal toxicity and acute pulmonary toxicity/

pathogenicity), non-irritating (primary dermal irritation), and not pathogenic (acute pulmonary 

toxicity/pathogenicity) (see section IV(B)(1)(a) on pages 8–10 and Table 5 in Appendix A).

b. Cumulative Effects from Substances with a Common Mechanism of Toxicity

Section 408(b)(2)(D)(v) of FFDCA requires that, when considering whether to establish, modify, 

or revoke a tolerance exemption, the EPA consider “available information concerning the 

cumulative effects of [a particular pesticide’s] . . . residues and other substances that have a 

common mechanism of toxicity.”

The EPA has not found Helicoverpa zea ABA Nucleopolyhedrovirus - U to share a common 

mechanism of toxicity with other substances, and Hz ABA NPV-U does not appear to produce a 

toxic metabolite against target pests. For the purposes of the tolerance action, therefore, the EPA 

has assumed that Hz ABA NPV-U does not have a common mechanism of toxicity with other 

substances. Thus, section 408(b)(2)(D)(v) of the FFDCA does not apply. For information 

regarding the EPA’s efforts to determine which chemicals have a common mechanism of 

toxicity and to evaluate the cumulative effects of such chemicals, see the EPA’s website at 

http://www.epa.gov/pesticides/cumulative.

c. Determination of Safety for the United States Population, Infants and Children

In considering the establishment of a tolerance or tolerance exemption for a pesticide chemical 

residue, FFDCA section 408(b)(2)(C) provides that the EPA shall assess the available 

information about consumption patterns among infants and children, special susceptibility of 

infants and children to pesticide chemical residues, and the cumulative effects on infants and 

children of the residues and other substances with a common mechanism of toxicity. In addition, 

FFDCA section 408(b)(2)(C) provides that the EPA shall apply an additional tenfold (10X) 

margin of exposure (safety) for infants and children in the case of threshold effects to account for 

prenatal and postnatal toxicity and the completeness of the database on toxicity and exposure 

unless the EPA determines that a different margin of exposure (safety) will be safe for infants 

and children. This additional margin of exposure (safety) is commonly referred to as the Food 

Quality Protection Act Safety Factor. In applying this provision, the EPA either retains the 

default value of 10X or uses a different additional safety factor when reliable data available to 

the EPA support the choice of a different factor.

Based on the acute toxicity and pathogenicity data/information discussed in section IV(B)(1)(a) 

(see pages 8–10) and Table 5 in Appendix A, the EPA concludes that there are no threshold 

effects of concern to infants, children, or adults when Helicoverpa zea ABA 

Nucleopolyhedrovirus - U is used as labeled in accordance with good agricultural practices. As a 

result, the EPA concludes that no additional margin of exposure (safety) is necessary to protect 

infants and children and that not adding any additional margin of exposure (safety) will be safe 

for infants and children.

Moreover, based on the same data/information and EPA analysis as presented directly above, the 

Agency is able to conclude that there is a reasonable certainty that no harm will result to the 

United States population, including infants and children, from aggregate exposure to the residues 

of Helicoverpa zea ABA Nucleopolyhedrovirus - U when it is used—as labeled and in 
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accordance with good agricultural practices—as an insecticide. Such exposure includes all 

anticipated dietary exposures and all other exposures for which there is reliable information. The 

EPA has arrived at this conclusion because, considered collectively, the data and information 

available on Hz ABA NPV-U do not demonstrate toxic and/ pathogenic potential to mammals, 

including infants and children. 

3. Occupational Exposure and Risk Characterization

The EPA does not expect handler exposure to Helicoverpa zea ABA Nucleopolyhedrovirus - U

to pose any undue risk. Regardless, appropriate personal protective equipment (PPE) and 

precautionary statements are required on pesticide product labels to mitigate any potential risks 

to pesticide handlers due to prolonged or numerous exposures. Handlers applying Heligen in 

agricultural settings must wear a dust/mist filtering respirator, waterproof gloves, long-sleeved 

shirt, long pants, socks and shoes. For future products containing this insect virus as an active 

ingredient, the EPA may require additional PPE (e.g., protective eyewear) on a product-specific 

basis.

4. Human Health Risk Characterization

The EPA considered human exposure to Helicoverpa zea ABA Nucleopolyhedrovirus - U in 

light of the standard for registration in FIFRA and the relevant safety factors in FFDCA. The 

Agency determined that no unreasonable adverse effects to the United States population in 

general, and to infants and children in particular, will result when Hz ABA NPV-U is used in 

accordance with EPA-approved labeling.

C. Environmental Assessment (40 CFR § 158.2150)

Data and other information (e.g., scientific literature) submitted and cited by the applicant to 

support requests to waive nontarget organism testing for Helicoverpa zea ABA 

Nucleopolyhedrovirus – U are sufficient to fulfill the relevant microbial pesticide data 

requirements and for risk assessment purposes. In addition, the applicant submitted one nonguideline study on nontarget insects that partially fulfilled this data requirement. Further testing 

of nontarget organisms at higher tier levels (i.e., Tiers II, III, and IV) is not required. The EPA

performed an environmental risk assessment based on data and other information provided by 

the applicant, and the Agency determined that adverse effects to nontarget organisms are not 

anticipated from the proposed pesticidal uses of Hz ABA NPV-U. Moreover, the EPA has made 

a “No Effect” determination for direct and indirect effects to federally listed threatened and 

endangered species and their designated critical habitats resulting from these same proposed 

pesticidal uses. 

For a summary of the generic data requirements described in sections IV(C)(1) (see pages 14–

15), refer to Table 6 in Appendix A.

1. Ecological Exposure and Risk Characterization 

The applicant evaluated the identity or similarity between NPV strains by extracting DNA, 

treating with restriction endonuclease and using electrophoresis to compare the gel patterns. The 

products, AgBiTech's Heligen and the previously registered product containing Hz NPV, 

Gemstar (EPA Registration Number 70051-45), were examined by light microscopy using 
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Giemsa stain and X I000 magnification under phase contrast. This process confirmed that the 

products contain a nucleopolyhedrovirus with occlusion bodies visually similar to each other. 

Researchers extracted the virus from both products, and they extracted DNA from the virus 

occlusion bodies. Then, this DNA was digested with restriction endonuclease (RENs) and the 

profile under electrophoresis on agarose was compared with that of NPV in Hz NPV (Gemstar). 

This analysis shows that the Hz ABA NPV-U in the AgBiTech product is almost identical to the 

Hz NPV found in Gemstar. Based on this, data previously submitted to support the registration 

of HzNPV (Gemstar) can be bridged to this new strain.

a. Terrestrial Animals and Plants

Wild Mammals (Harmonized Guideline 885.4150, MRID No. 489417-06), Birds (Harmonized 

Guideline 885.4100), Nontarget Insects (Harmonized Guideline 885.4340, MRID No. 489418-

10), Honey Bees (Harmonized Guideline 885.4380, MRID No. 489418-09) and Plants 

(Harmonized Guideline 885.4300): 

The proposed applications—ground and aerial sprays and sprinkler chemigation –would deposit 

Hz ABA NPV-U on foliar and other above ground plant surfaces. EPA expects deposition onto 

these surfaces to occur within the intended area of treatment, as well as surrounding areas. 

Persistence of Hz ABA NPV-U in the environment, particularly in soil, is possible as long as 

conditions are favorable, but it does not pose risks to nontarget species because of its specificity. 

Testing on birds, wild mammals, nontarget insects, and honey bees were not required because 

the applicant instead submitted acceptable scientific rationale and/or cited previous studies. The 

EPA additionally determined that, since Nz ABA NPV-U is not taxonomically related to known 

plant pathogens, adverse effects to nontarget plants are not expected. Baculoviruses such as Hz 

NPV have a history of safe use as microbial pesticides, and they generally have narrow host 

ranges with no adverse effects on nontarget organisms (OECD 2002). Typically, NPVs only 

infect the genus or perhaps family from which they were originally isolated. Searches of public 

literature did not indicate reports of adverse effects of Hz NPV to these nontarget organisms. 

An additional bioassay with a nontarget insect, Spodoptera frugiperda, was submitted (MRID 

48941809). This study tested the toxicity of Hz ABA NPV-U to this insect, and results indicated 

the potential for some effects in this nontarget lepidopteran. However, effects to S. frugiperda

would not be expected at field application rates, and sensitivity is expected to be less in species 

that are not as closely related to the target pest. See USEPA (2013d). Based on the information 

submitted and cited by the applicant, adverse effects to terrestrial nontarget organisms are not 

expected to result from the proposed pesticidal uses of Hz ABA NPV-U.

b. Aquatic Animals and Plants

Freshwater Fish (Harmonized Guideline 885.4200), Freshwater Invertebrates (Harmonized 

Guideline 885.4240), Estuarine/Marine Fish and Invertebrates (Harmonized Guideline 

885.4280) (MRID No. 489418-10):

The EPA determined that freshwater and estuarine fish and invertebrates are not likely to be 

exposed to Hz ABA NPV-U from runoff and drift into aquatic areas and that Hz ABA NPV-U is 

not pathogenic to aquatic organisms. The applicant submitted acceptable scientific rationale 

and/or cited previous studies. Baculoviruses such as Hz NPV have a history of safe use as 

microbial pesticides, and they generally have narrow host ranges with no adverse effects on 
Helicoverpa zea Nucleopolyhedrovirus Page 16 of 27

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nontarget organisms (OECD 2002). Typically, NPVs only infect the genus or perhaps family 

from which they were originally isolated. Searches of public literature did not indicate reports of 

adverse effects of Hz NPV to these nontarget organisms. 

Based on the information submitted and cited by the applicant, adverse effects to aquatic 

nontarget organisms are not expected to result from the proposed pesticidal uses of Hz ABA 

NPV-U.

2. Environmental Fate Data 

As the data and information provided are sufficient to fulfill the Tier I nontarget organism data 

requirements and allow for nontarget organism risk assessment for Helicoverpa zea ABA 

Nucleopolyhedrovirus-U, further testing at higher levels (Tiers II, III, and IV) is not required. 

3. Threatened and Endangered Species Assessment

Since the EPA has determined that no effects are anticipated for any nontarget species exposed 

to Helicoverpa zea ABA Nucleopolyhedrovirus - U as a result of the proposed labeled uses, 

effects to federally listed threatened and endangered species and their designated critical habitats 

also are not expected. Therefore, a “No Effect” determination is made for direct and indirect 

effects to listed species and their designated critical habitats resulting from the proposed uses of

Hz ABA NPV-U.

V. ENVIRONMENTAL JUSTICE

The EPA seeks to achieve environmental justice—the fair treatment and meaningful involvement 

of all people regardless of race, color, national origin, or income—with respect to the 

development, implementation, and enforcement of environmental laws, regulations, and policies. 

Fair treatment means that no group of people, including racial, ethnic, or socioeconomic groups,

should bear a disproportionate share of the negative environmental consequences resulting from 

industrial, municipal, and commercial operations or the execution of federal, state, local, and 

tribal environmental programs and policies. Meaningful involvement means that (1) potentially 

affected community residents have an appropriate opportunity to participate in decisions about a 

proposed activity that will affect their environment and/or health; (2) the public’s contribution 

can influence the regulatory agency’s decision; (3) the concerns of all participants involved will 

be considered in the decision-making process; and (4) the decision-makers seek out and facilitate 

the involvement of those potentially affected. The EPA has this goal for all communities and 

persons across the United States. 

To help address potential environmental justice issues, the EPA seeks information on any groups 

or segments of the population who, as a result of their location, cultural practices, or other 

factors, may have atypical, unusually high exposure to Helicoverpa zea ABA 

Nucleopolyhedrovirus - U compared to the general population. Please comment if you are aware 

of any subpopulations that may have atypical, unusually high exposure compared to the general 

population.

For additional information regarding environmental justice issues, please visit the EPA’s web 

site at http://www.epa.gov/compliance/environmentaljustice/index.html.
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VI. RISK MANAGEMENT DECISION

Section 3(c)(5) of FIFRA permits for the registration of a pesticide provided that all the 

following determinations are made:

(1) Its composition is such as to warrant the proposed claims for it;

(2) Its labeling and other material required to be submitted comply with the

 requirements of FIFRA;

(3) It will perform its intended function without unreasonable adverse effects on 

 the environment; AND

(4) When used in accordance with widespread and commonly recognized 

 practice, it will not generally cause unreasonable adverse effects on the 

 environment.

To satisfy criterion 1, the Helicoverpa zea ABA Nucleopolyhedrovirus - U products, MP and EP,

have well-known properties. The EPA has no knowledge that would contradict the claims made 

on the labels, and the pesticide products are not expected to cause unreasonable adverse effects

on the environment when used according to label instructions. Criterion 2 is satisfied by the 

current product labels, as well as the data and information presented in this document. The 

Agency believes that the Hz ABA NPV-U pesticide products will not cause any unreasonable 

adverse effects on the environment. In fact, the EP, Heligen, is likely to provide protection 

against many infestations, satisfying criterion 3. Criterion 4 is satisfied in that the Hz ABA NPVU pesticide products are not expected to cause unreasonable adverse effects when used according 

to label instructions. Therefore, both the MP and the EP, Heligen, which contains Hz ABA NPVU as a new active ingredient, are eligible for registration under FIFRA section 3(c)(5) for the

labeled uses. 

VII. ACTIONS REQUIRED OF THE REGISTRANT

A. Final Printed Labeling

Before the proposed manufacturing-use product, Helicoverpa zea ABA Nucleopolyhedrovirus -

U, and the proposed end-use product, Heligen, may be released for shipment, the applicant is 

required to provide appropriate final printed labeling to the EPA.

B. Terms of Registration

The EPA is not requiring additional information on the registration of these two products.

C. Reporting of Adverse Effects and Hypersensitivity Incidents

Notwithstanding the information stated in the previous sections, it should be clearly understood 

that certain specific data must be reported to the EPA as a requirement for maintaining the 

federal registration for a pesticide product. Reports of all incidents of adverse effects to the 

environment must be submitted to the EPA under the provisions stated in FIFRA section 6(a)(2). 

Additionally, all incidents of hypersensitivity (including both suspected and confirmed incidents) 

must be reported to the EPA under the provisions of 40 CFR § 158.2140(d).
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VIII. GLOSSARY OF ACRONYMS AND ABBREVIATIONS

BRAD Biopesticides Registration Action Document

CFR Code of Federal Regulations

cSt centistokes

EDSP Endocrine Disruptor Screening Program

EP end-use pesticide product

EPA United States Environmental Protection Agency (the “Agency”)

FFDCA Federal Food, Drug, and Cosmetic Act

FIFRA Federal Insecticide, Fungicide, and Rodenticide Act

FR Federal Register

g/mL grams per milliliter

Hz NPV Helicoverpa zea Nucleopolyhedrovirus product

lb/gal pounds per gallon

LD50 median lethal dose. A statistically derived single dose that can be expected 

to cause death in 50% of the test animals when administered by the route 

indicated (oral, dermal, or inhalation). It is expressed as a weight of 

substance per unit weight of animal (e.g., mg/kg).

mg/kg milligrams per kilogram

MP manufacturing-use pesticide product

MRID No. Master Record Identification Number

NIOSH National Institute for Occupational Safety and Health

OPP Office of Pesticide Programs

PC Code Pesticide Chemical Code

PP Pesticide Petition

PPE personal protective equipment

TGAI technical grade of the active ingredient

U.S. United States
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IX. BIBLIOGRAPHY

A. Studies Submitted to Support the Active Ingredient, Helicoverpa zea ABA Nucleopolyhedrovirus - U, and Pesticide Product Registrations for the MP and the EP, Heligen.

MRID No. Study Information

48941700 AgBiTech Pty Ltd. (2012) Submission of Product Chemistry and Toxicity Data in 

Support of the Application for Registration of Heligen. Transmittal of 7 Studies.

48941701 MacIntosh, S. (2012) Product Identity, Manufacturing Process, Product 

Characterization: Viscosity & Discussion of Unintentional Ingredients: for Heligen 

(EP). Unpublished study prepared by MacIntosh & Associates, Inc. 17p.

48941702 MacIntosh, S. (2012) Analysis of Samples: for Heligen (EP). Unpublished study 

prepared by MacIntosh & Associates, Inc. 15p.

48941703 Hawes, A. (2012) Certification of Limits: for Heligen (EP). Unpublished study 

prepared by AgBiTech Pty Ltd. 8p.

48941704 Hawes, A. (2006) Storage Stability & Corrosion Characteristics for Heligen (EP). 

Project Number: 06/001/2. Unpublished study prepared by Queensland Dept. of 

Primary Industries, Sonic Food and Water Testing and AgBiTech Pty Ltd. 18p.

48941705 Hawes, A. (2012) Hypersensitive Incidents: Helifen (EP). Unpublished study 

prepared by AGBITECH PTY LTD. 6p.

48941706 MacIntosh, S. (2012) Statisfaction of Data Requirements with Public Literature 

and/or Scientific Rationale: Acute Oral Toxicity: Acute Dermal Toxicity: Acute 

Inhalation Toxicity: Acute Eye Irritation Toxicity: Acute Dermal Irritation Toxicity: 

for Heligen. Unpublished study prepared by MacIntosh & Associates, Inc. 14p.

48941707 MacIntosh, S. (2012) Satisfaction of Data Requirements with Public Literature and/or 

Scientific Rationale: Freshwater Fish Toxicity/Pathogenicity: Freshwater Invertebrate 

Toxicity/Pathogenicity: for Heligen (EP). Unpublished study prepared by MacIntosh 

& Associates, Inc. 9p.

48941800 AgBiTech Pty Ltd. (2012) Submission of Product Chemistry and Toxicity Data in 

Support of the Application for Registration of Helicoverpa zea Nucleopolyhedrovirus. 

Transmittal of 10 Studies.

48941801 MacIntosh, S. (2012) Product Identity, Manufacturing Process & Discussion of 

Unintentional Ingredients: for Helicoverpa zea Nucleopolyhedrovirus. Unpublished 

study prepared by MacIntosh & Associates, Inc. 23p.

48941802 MacIntosh, S. (2012) Analysis of Samples: for Helicoverpa zea 

Nucleopolyhedrovirus (TGAI/MP). Unpublished study prepared by MacIntosh & 

Associates, Inc. 16p.
Helicoverpa zea Nucleopolyhedrovirus Page 20 of 27

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MRID No. Study Information

48941803 Hawes, A. (2012) Certification of Limits: for Helicoverpa zea Nucleopolyhedrovirus. 

Unpublished study prepared by AgBiTech Pty Ltd. 7p.

48941804 Hawes, A. (2012) Product Chemistry: for Helicoverpa zea Nucleopolyhedrovirus.

 Unpublished study prepared by AgBiTech Pty Ltd. 7p.

48941805 MacIntosh, S. (2012) Satisfaction of Data Requirements with Public Literature

 and/orScientific Rationale: Storage Stability and Corrosion Characteristics for

 Helicoverpa zea Nucleopolyhedrovirus (TGAI/MP). Unpublished study prepared by 

 MacIntosh & Associates, Inc. 15p.

48941806 Hawes, A. (2012) Hypersensitive Incidents: Helicoverpa zea Nucleopolyhedrovirus 

(TGAI/MP). Unpublished study prepared by AgBiTech Pty Ltd. 6p.

48941807 Pavloff, N. (2012) Cell Culture Tests for a Helicoverpa zea Nucleopolyhedrovirus. 

Project Number: 173/4P/1R/CB/04/15/11. Unpublished study. 22p.

48941808 MacIntosh, S. (2012) Satisfaction of Data Requirements with Public Literature and/or 

Scientific Rationale: for Helicoverpa zea Nucleopolyhedrovirus. Unpublished study 

prepared by MacIntosh & Associates, Inc. 18p.

48941809 Popham, H. (2012) Non-Target Insect Bioassay: for Helicoverpa zea Nucleopolyhedrovirus. Project Number: 12/016. Unpublished study prepared by USDA-ARS 

BCIRL. 13p.

48941810 MacIntosh, S. (2012) Satisfaction of Data Requirements with Public Literature and/or 

Scientific Rationale: for Helicoverpa zea nucleopolyhedrovirus (TGAI/MP). 

Unpublished study prepared by MacIntosh & Associates, Inc. 14p.

49011400 AgBiTech Pty, Ltd. (2012) Submission of Product Chemistry and Toxicity Data in

Support of the Applications for Registration of Helicoverpa zea Nucleopolyhedrovirus and Heligen. Transmittal of 5 Studies.

49011401 MacIntosh, S. (2012) Supplemental Information Product Identity: for Helicoverpa zea 

Nucleopolyhedrovirus Supplemental to MRID No 489418-01. Unpublished study 

prepared by MacIntosh & Associates, Inc. 22p.

49011402 MacIntosh, S. (2012) Satisfaction of Data Requirements with Public Literature and/or 

Scientific Rationale for HzNPV for Toxicology Guideline Studies Supplement to:

MRID No 489418-08. Unpublished study by MacIntosh & Associates, Inc.13p.

49011403 MacIntosh, S. (2012) Satisfaction of Data Requirements with Public Literature and/or 

Scientific Rationale for HzNPV for Non-target Organism Guideline Studies 

Supplement to: MRID No 489418-10. Unpublished study prepared by MacIntosh & 

Associates, Inc. 13p.
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MRID No. Study Information

49011404 MacIntosh, S. (2012) Satisfaction of Data Requirements with Public Literature and/or 

Scientific Rationale for Heligen for Toxicology Guideline Studies Supplement to: 

MRID No 489417-06. Unpublished study prepared by MacIntosh & Associates, Inc. 

13p.

49011405 MacIntosh, S. (2012) Publications in Support of Registration Applications for

Helicoverpa zea Nucleopolyhedrovirus and Heligen. Unpublished study prepared by 

MacIntosh & Associates, Inc. 322p.

49020400 AgBiTech Pty, Ltd. (2012) Submission of Product Chemistry Data in Support of the 

Applications for Registration of Helicoverpa zea Nucleopolyhedrovirus and Heligen. 

Transmittal of 1 Study.

49020401 Popham, H.; Hawes, A. (2012) Supplemental Information: Product Identity: For 

Helicoverpa zea Nucleopolyhedrovirus. Unpublished study prepared by Agbitech, Ltd 

and U.S. Dept. of Agriculture. 35p.

B. Environmental Protection Agency Risk Assessment Memoranda

U.S. EPA. 2013a. Review of Product Chemistry, Manufacturing Process, and Acute Toxicity 

Studies of the TGAI Helicoverpa zea Nucleopolyhedrovirus (HZ NPV) for section 3 

product registration (EPA Reg. no. 87978-R). Memorandum from Ibrahim S. Barsoum, 

Ph.D. and John L. Kough, Ph.D. to Michael Glikes, dated June 4, 2013.

U.S. EPA. 2013b. Review of Product Chemistry, Manufacturing Process, and Acute Toxicity 

Studies of the EP Heligen containing Helicoverpa zea Nucleopolyhedrovirus (HZ NPV) 

TGAI for section 3 product registration (EPA Reg. no. 87978-E). Memorandum from 

Ibrahim S. Barsoum, Ph.D. and John L. Kough, Ph.D. to Michael Glikes, dated June 12, 

2013.

U.S. EPA. 2013c. Review of the registrant’s response to the identification and concentration of 

the surfactant used in the manufacturing process of the EP Heligen containing 

Helicoverpa zea Nucleopolyhedrovirus (HZ NPV) (EPA Reg. no. 87978-E). 

Memorandum from Ibrahim S. Barsoum, Ph.D. and John L. Kough, Ph.D. to Michael 

Glikes, dated June 10, 2013.

U.S. EPA. 2013d. Environmental Risk Assessment of Polyhedral occlusion bodies (OBs) of the 

nuclear polyhedrosis virus of Helicoverpa zea for FIFRA Section 3 Registrations of 

Helicoverpa zea Nucleopolyhedrovirus Manufacturing Use product (EPA File Symbol 

87978-R) and Heligen End Use Product (EPA File Symbol 87978-R) for Control of 

Helicoverpa zea and Heliothis virescens in Food and Nonfood Crops; PC Code: 107300; 

Decision No. 469835 and 469836; DP Barcodes: 408136 and 408138; Submission Nos. 

928090 and 928232; MRIDs 48941809, 48941810, 49011403, and 48941707.

U.S. EPA. Registration Eligibility Decision (RED) Facts: Heliothis Zea NPV, January 1991.
Helicoverpa zea Nucleopolyhedrovirus Page 22 of 27

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C. Other References

Annals of the Entomological Society of America. 2004. A New Synonym of Helicoverpa zea

(Boddie) and Differentiation of Adult Males of H. zea and H. armigera (Hubner). Vol. 

97, no. 6.

Centers for Disease Control and Prevention. 2009. Drinking Water - Water Treatment. Available 

from http://www.cdc.gov/healthywater/drinking/public/water_treatment.html.

DeFelice K, Wollenhaupt N, Buchholz D. 1993. Aquifers and Soil Filter Effect. Available from 

http://extension.missouri.edu/p/WQ24.

U.S. EPA. 2004. Primer for Municipal Wastewater Treatment Systems. EPA 832-R-04-001. 

Available from http://www.epa.gov/npdes/pubs/primer.pdf.
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APPENDIX A. MICROBIAL PESTICIDES DATA REQUIREMENTS 

(40 CFR PART 158 – SUBPART V)

TABLE 1. Product Analysis Data Requirements for the Active Ingredient (TGAI), Helicoverpa zea AB Nucleopolyhedroviru- Us

(40 CFR § 158.2120)

Harmonized 

Guideline 

Number

Data Requirement Results MRID No.

885.1100 Product Identity Submitted data fulfill the requirement for product identity and composition. 48941801

885.1200 Manufacturing 

Process Submitted data fulfill the requirement for manufacturing process. 48941801

N/A

Deposition of a 

Sample in a 

Nationally 

Recognized Culture 

Collection

Helicoverpa zea ABA Nucleopolyhedrovirus - U is on deposit with the ATCC.

48941801

885.1300

Discussion of 

Formation of 

Unintentional 

Ingredients

Submitted data fulfill the requirement for preventing and detecting any 

microbial contaminants. 48941801

885.1400 Analysis of Samples Submitted data fulfill the requirement for analysis of samples. 48941802

885.1500

Certification of 

Limits Submitted data fulfill the requirement for manufacturing process. 48941803

TABLE 2. Product Analysis Data Requirements for the End-Use Pesticide Product (EP), Heligen

(40 CFR § 158.2120)

Harmonized

Guideline 

Number

Data Requirement Results MRID No.

885.1100 Product Identity Submitted data fulfill the requirement for product identity. 48941701

885.1200 Manufacturing Process Submitted data fulfill the requirement for manufacturing 

process.

48941701

N/A

Deposition of a Sample in a 

Nationally Recognized 

Culture Collection

Submitted data fulfill the requirement for deposition of a sample 

in a culture collection. N/A

885.1300 Discussion of Formation of 

Unintentional Ingredients

Submitted data fulfill the requirement for preventing and 

detecting any microbial contaminants. 48941701

885.1400 Analysis of Samples Submitted data fulfill the requirement for analysis of samples 

for purposes of FIFRA section 3(c)(5) registration. 

48941702

885.1500 Certification of Limits Limits listed on the confidential statement of formula are 

adequate/acceptable. 48941703
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TABLE 3. Physical and Chemical Characteristics for the Technical Grade of the Active Ingredient (TGAI), Helicoverpa zea ABA

Nucleopolyhedrovirus - U (40 CFR § 158.2120)

Harmonized 

Guideline Number Data Requirement Results MRID No.

830.6302 Color Grey-brown translucent liquid

830.6303 Physical State Liquid 48941804

830.6304 Odor Earthy

830.6313

Stability to Normal and 

Elevated

Temperatures, Metals,

and Metal Ions

N/A N/A

830.6317 Storage Stability Submitted data fulfill the requirement for storage stability. 48941805

830.6319 Miscibility Non emulsifiable liquid. N/A

830.6320 Corrosion 

Characteristics

Submitted data fulfill the requirement for corrosion 

characteristics. 48941805

830.7000 pH 5.0 – 6.0 48941804

830.7100 Viscosity N/A (not a homogenous solution) N/A

830.7300

Density/Relative 

Density/Bulk Density 

(Specific Gravity)

1.15 – 1.26 kg/L 48941804

TABLE 4. Physical and Chemical Characteristics for the End-Use Pesticide Product (EP), Heligen (40 CFR § 158.2120)

Harmonized

Guideline Number Data Requirement Results MRID No.

830.63021 Color Grey-brown translucent liquid 48941704

830.63031Physical State Liquid 48941704

830.63041 Odor Earthy 48941704

830.63131

Stability to Normal and 

Elevated Temperatures, 

Metals, and Metal Ions

N/A N/A

830.6317 Storage Stability Submitted data fulfill the requirement for storage 

stability. 48941704

830.6319 Miscibility

Not required because the end-use pesticide product, 

Serenade Soil DPZ, is not an emulsifiable liquid form of 

microbial pesticides (refer to test note #2 of 40 CFR § 

158.2120(d)).

N/A

830.6320 Corrosion Characteristics Submitted data fulfill the requirement for corrosion 

characteristics 48941704

830.7000 pH 5.0 – 6.0 48941704

830.7100 Viscosity 6.05 48941704

830.7300

Density/Relative 

Density/Bulk Density 

(Specific Gravity)

1.15 – 1.26 kg/L 48941704

1 According to 40 CFR § 158.2120, these data are only required for the technical grade of the active ingredient. Since AgBiTech 

LTD included this information with its application for Heligen, it is summarized appropriately in this table.
Helicoverpa zea Nucleopolyhedrovirus Page 25 of 27

Biopesticides Registration Action Document

TABLE 5. Toxicology Data Requirements for the Active Ingredient Helicoverpa zea ABA Nucleopolyhedrovirus - U

and the End Use Pesticide Product Heligen (40 CFR § 158.2140)

Harmonized 

Guideline 

Number

Data Requirement Results MRID No.

Tier I

885.3050 Acute Oral 

Toxicity/Pathogenicity

Waived based on extensive list of toxicology studies on other strains of 

NPV.

Classification: Acceptable

TOXICITY CATEGORY IV

48941706

885.3150 Acute Pulmonary 

Toxicity/Pathogenicity

Waived based on extensive list of toxicology studies on other strains of 

NPV.

Classification: Acceptable 48941706

885.3200

Acute Injection 

Toxicity/

Pathogenicity

Waived based on extensive list of toxicology studies on other strains of 

NPV.

Classification: Acceptable

48941706

885.3200

Acute Injection 

Toxicity/Pathogenicity 

(Intravenous)

Waived based on extensive list of toxicology studies on other strains of 

NPV.

Classification: Acceptable

48941706

885.3400 Hypersensitivity

Incidents

The applicant reported that no hypersensitivity incidents, including 

immediate-type or delayed-type reactions of humans and domestic animals, 

occurred during research, development, or testing of the TGAI. Any future 

hypersensitivity incidents must be reported to EPA (refer to test note #3 of 

40 CFR § 158.2140(d)). 

48941806

885.3500 Cell Culture

No cytopathic effects or replication of Hz NPV exposed to any of the 

mammalian cells were observed.

Classification: Acceptable

48941807

870.1100 Acute Oral Toxicity

Waived based on extensive list of toxicology studies on other strains of 

NPV.

Classification: Acceptable

TOXICITY CATEGORY IV

48941706

870.1200 Acute Dermal Toxicity

Waived based on extensive list of toxicology studies on other strains of 

NPV.

Classification: Acceptable

TOXICITY CATEGORY IV

48941706

870.1300 Acute Inhalation 

Toxicity

Waived based on extensive list of toxicology studies on other strains of 

NPV.

Classification: Acceptable

TOXICITY CATEGORY IV

48941706

870.2400 Acute Eye Irritation

Waived based on extensive list of toxicology studies on other strains of 

NPV.

Classification: Acceptable

TOXICITY CATEGORY IV

48941706

870.2500 Primary Dermal 

Irritation

Waived based on extensive list of toxicology studies on other strains of 

NPV.

Classification: Acceptable

TOXICITY CATEGORY IV

48941706

NA NA Restriction Endonuclease Analysis

Classification: Acceptable 49011402

Tiers II and III

Not required for Helicoverpa zea ABA Nucleopolyhedrovirus - U based on the lack of acute toxicity/pathogenicity in the Tier I studies.
Helicoverpa zea Nucleopolyhedrovirus Page 26 of 27

Biopesticides Registration Action Document

TABLE 6. Nontarget Organism Toxicity and Environmental Fate Data Requirements for the Technical Grade of the Active 

Ingredient (TGAI), Helicoverpa zea ABA Nucleopolyhedrovirus-U (40 CFR § 158.2150)

Harmonized Guideline 

Number

Data Requirement Results MRID No.

Tier I

885.4050 Avian Oral Toxicity

Scientific rationale, in addition to citation of 

MRIDs 000814446 and 00081477, provide

sufficient information to determine that 

toxicity/pathogenicity to avian wildlife is not 

expected from the proposed pesticidal uses.

Classification: Acceptable

48941810

885.4100 Avian Inhalation 

Toxicity/Pathogenicity

Not required as the nature of the microbial 

pesticide does not indicate potential pathogenicity 

to birds or relatedness to any known bird 

pathogens (refer to test note #3 of 40 CFR § 

158.2150(e)).

N/A

885.4150 Wild Mammal 

Toxicity/Pathogenicity

Scientific rationale indicates that adverse effects 

to mammals would not be expected.

Classification: Acceptable for wild mammal 

risk assessment

48941810

885.4200 Freshwater Fish 

Toxicity/Pathogenicity

Scientific rationale, in addition to citation of 

MRIDs 00081444 and 00081445, provide 

sufficient information to determine that 

toxicity/pathogenicity to freshwater fish and 

invertebrates is not expected from the pesticide.

Classification: Acceptable

48941810

885.4240 Freshwater Invertebrate 

Toxicity/Pathogenicity

885.4280 Estuarine/Marine Fish and 

Invertebrate Testing

Not required because of the established 

toxicity/pathogenicity profile, narrow host range, 

and history of Hz NPV use without apparent 

adverse effects to estuarine/marine animals.

N/A

885.4300 Nontarget Plant Testing

Not required as the microbial pesticide is not 

taxonomically related to any known plant 

pathogens (refer to test note #7 of 40 CFR § 

158.2150(e)).

N/A

885.4340 Nontarget Insect Testing

Scientific rationale, in addition to citation of 

MRID 00049121, provides sufficient information 

to determine that toxicity/pathogenicity to honey 

bees and nontarget insects is not expected as a 

result of the proposed pesticidal uses. A 

nonguideline study also provided information on 

the potential for effects to Spodoptera frugiperda.

Classification: Acceptable

48941809 

48941810

885.4380 Honey Bee Testing

Tiers II, III, and IV

Not required for Helicoverpa zea ABA Nucleopolyhedrovirus - U based on the acceptability of the data and other information provided 

for Tier I.
Helicoverpa zea Nucleopolyhedrovirus Page 27 of 27

Biopesticides Registration Action Document

APPENDIX B. PESTICIDE PRODUCTS

EPA File 

Symbol

Registration 

Name

Percenta

ge 

Active 

Ingredie

nt

Formulation 

Type

Use Site(s) Method(s) of 

Application

Application Rate(s) Target Pest

87978-R

Helicoverpa 

zea ABA 

Nucleopolyhedrovirus - U

74.5% Manufacturinguse product N/A N/A N/A

Cotton Bollworm 

(Helicoverpa zea) and 

Tobacco budworm 

(Heliothis virescens)

87978-E Heligen 32% End-use 

product

Agricultural

Ground application

Aerial

Chemigation

0.7 – 2.4 fl. oz./ acre 

depending on the crop

Cotton Bollworm 

(Helicoverpa zea) and 

Tobacco budworm 

(Heliothis virescens)